Development Of A Naive Human Fab Antibody Library To Generate Monoclonal Antibodies Against Bmsxp Recombinant Antigen
| dc.contributor.author | Omar, Noorsharmimi | |
| dc.date.accessioned | 2016-07-25T07:48:41Z | |
| dc.date.available | 2016-07-25T07:48:41Z | |
| dc.date.issued | 2015-09 | |
| dc.description.abstract | Antibodies are glycoproteins produced by B-cells against molecules foreign to the body. This study focused on the production of antigen specific human The Fab region consists of one constant and one variable domain which are both located at the heavy chain and the light chain. An optimized protocol for the production of a recombinant Fab library of unimmunized donors from 3 major ethnic groups (Chinese, Malay and Indian) in Malaysia was carried out. We used peripheral blood lymphocytes total ribonucleic acids (RNA) from 90 donors to obtain variable genes (V-gene) specific cDNA. The Fab-encoding regions of IgM repertoire were individually amplified by polymerase chain reaction (PCR) using a diverse set of primers corresponding to the different variable region of the heavy (μ) and the light chains (к,λ). The diversity of the library is created via random combinatorial mixing of all heavy and light V-genes. A highly diverse naïve library is useful for the selection of antibodies against a panel of antigens. Phage selection was carried out against BmSXP using two different panning strategies to understand the efficiency of Fab library panning with different strategies. BmSXP is a recombinant antigen of Wuchereria Bancrofti which is specific for Bancroftian lymphatic filarial disease and is used for diagnostic of lymphatic filarial infection. The performance of both panning strategies suggests the difficulty for Fab library panning with an automated process as only the conventional panning method was able to isolate positive binders. Two monoclonal Fab antibodies against BmSXP were identified and confirmed. Although difficult, the production of a Fab naïve library is useful for the generation of monoclonal Fab antibodies for downstream applications such as diagnostics. Phage display panning of a naïve Fab library is able to select monoclonal antibodies against the target antigens efficiently. This provides an attractive solution for the generation of recombinant human antibodies. | en_US |
| dc.identifier.uri | http://hdl.handle.net/123456789/2302 | |
| dc.title | Development Of A Naive Human Fab Antibody Library To Generate Monoclonal Antibodies Against Bmsxp Recombinant Antigen | en_US |
| dc.type | Thesis | en_US |
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